Two-Dimensional Crystallography of TFIIB– and IIE–RNA Polymerase II Complexes: Implications for Start Site Selection and Initiation Complex Formation

AbstractTranscription factors IIB (TFIIB) and IIE (TFIIE) bound to RNA polymerase II have been revealed by electron crystallography in projection at 15.7 Å resolution. The results lead to simple hypotheses for the roles of these factors in the initiation of transcription. TFIIB is suggested to define the distance from TATA box to transcription start site by bringing TATA DNA in contact with polymerase at that distance from the active center of the enzyme. TFIIE is suggested to participate in a key conformational switch occurring at the active center upon polymerase–DNA interaction

The Challenge of Fetal Cardiac MRI Reconstruction Using Deep Learning

Dynamic free-breathing fetal cardiac MRI is one of the most challenging modalities, which requires high temporal and spatial resolution to depict rapid changes in a small fetal heart. The ability of deep learning methods to recover undersampled data could help to optimise the kt-SENSE acquisition strategy and improve non-gated kt-SENSE reconstruction quality. In this work, we explore supervised deep learning networks for reconstruction of kt-SENSE style acquired data using an extensive in vivo dataset. Having access to fully-sampled low-resolution multi-coil fetal cardiac MRI, we study the performance of the networks to recover fully-sampled data from undersampled data. We consider model architectures together with training strategies taking into account their application in the real clinical setup used to collect the dataset to enable networks to recover prospectively undersampled data. We explore a set of modifications to form a baseline performance evaluation for dynamic fetal cardiac MRI on real data. We systematically evaluate the models on coil-combined data to reveal the effect of the suggested changes to the architecture in the context of fetal heart properties. We show that the best-performers recover a detailed depiction of the maternal anatomy on a large scale, but the dynamic properties of the fetal heart are under-represented. Training directly on multi-coil data improves the performance of the models, allows their prospective application to undersampled data and makes them outperform CTFNet introduced for adult cardiac cine MRI. However, these models deliver similar qualitative performances recovering the maternal body very well but underestimating the dynamic properties of fetal heart. This dynamic feature of fast change of fetal heart that is highly localised suggests both more targeted training and evaluation methods might be needed for fetal heart application

Diagnostic Potential of the NMDA Receptor Peptide Assay for Acute Ischemic Stroke

Background The acute assessment of patients with suspected ischemic stroke remains challenging. The use of brain biomarker assays may improve the early diagnosis of ischemic stroke. The main goal of the study was to evaluate whether the NR2 peptide, a product of the proteolytic degradation of N-methyl-D-aspartate (NMDA) receptors, can differentiate acute ischemic stroke (IS) from stroke mimics and persons with vascular risk factors/healthy controls. A possible correlation between biomarker values and lesion sizes was investigated as the secondary objective. Methods and Findings A total of 192 patients with suspected stroke who presented within 72 h of symptom onset were prospectively enrolled. The final diagnosis was determined based on clinical observations and radiological findings. Additionally gender- and age-matched healthy controls (n = 52) and persons with controlled vascular risk factors (n = 48) were recruited to compare NR2 peptide levels. Blinded plasma was assayed by rapid magnetic particles (MP) ELISA for NR2 peptide within 30 min and results for different groups compared using univariate and multivariate statistical analyses. There was a clinical diagnosis of IS in 101 of 192 (53%) and non-stroke in 91 (47%) subjects. The non-stroke group included presented with acute stroke symptoms who had no stroke (n = 71) and stroke mimics (n = 20). The highest NR2 peptide elevations where found in patients with IS that peaked at 12 h following symptom onset. When the biomarker cut off was set at 1.0 ug/L, this resulted in a sensitivity of 92% and a specificity of 96% to detect IS. A moderate correlation (rs = 0.73) between NR2 peptide values and acute ischemic cortical lesions (\u3c200 \u3emL) was found. Conclusions This study suggests that the NR2 peptide may be a brain specific biomarker to diagnose acute IS and may allow the differentiation of IS from stroke mimics and controls. Additional larger scale clinical validation studies are required

Comparison of retinal vasodilator and constrictor responses in type 2 diabetes

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/93551/1/j.1755-3768.2012.02445.x.pd

Nanoproteomic analysis of ischemia-dependent changes in signaling protein phosphorylation in colorectal normal and cancer tissue

Additional file 1: Table S1. Clinical data for the 20 patients analyzed in the study. Presents patient clinical data including tumor stage and grade

Impaired retinal vasodilator responses in prediabetes and type 2 diabetes

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/99677/1/aos12129.pd

Dopamine D1 vs D5 receptor-dependent induction of seizures in relation to DARPP-32, ERK1/2 and GluR1-AMPA signalling.

Recent reports have shown that the selective dopamine D(1)-like agonist SKF 83822 [which stimulates adenylate cyclase, but not phospholipase C] induces prominent behavioral seizures in mice, whereas its benzazepine congener SKF 83959 [which stimulates phospholipase C, but not adenylate cyclase] does not. To investigate the relative involvement of D(1) vs D(5) receptors in mediating seizures, ethological behavioral topography and cortical EEGs were recorded in D(1), D(5) and DARPP-32 knockout mice in response to a convulsant dose of SKF 83822. SKF 83822-induced behavioral and EEG seizures were gene dose-dependently abolished in D(1) knockouts. In both heterozygous and homozygous D(5) knockouts, the latency to first seizure was significantly increased and total EEG seizures were reduced relative to wild-types. The majority (60%) of homozygous DARPP-32 knockouts did not have seizures; of those having seizures (40%), the latency to first seizure was significantly increased and the number of high amplitude, high frequency polyspike EEG events was reduced. In addition, immunoblotting was performed to investigate downstream intracellular signalling mechanisms at D(1)-like receptors following challenge with SKF 83822 and SKF 83959. In wild-types administered SKF 83822, levels of ERK1/2 and GluR1 AMPA receptor phosphorylation increased two-fold in both the striatum and hippocampus; in striatal slices DARPP-32 phosphorylation at Thr34 increased five-fold relative to vehicle-treated controls. These findings indicate that D(1), and to a lesser extent D(5), receptor coupling to DARPP-32, ERK1/2 and glutamatergic signalling is involved in mediating the convulsant effects of SKF 83822